Venting of carbon dioxide-rich fluid and hydrate formation in mid-okinawa trough backarc basin.

Carbon dioxide-rich fluid bubbles, containing approximately 86 percent CO(2), 3 percent H(2)S, and 11 percent residual gas (CH(4) + H(2)), were observed to emerge from the sea floor at 1335- to 1550-m depth in the JADE hydrothermal field, mid-Okinawa Trough. Upon contact with seawater at 3.8 degrees C, gas hydrate immediately formed on the surface of the bubbles and these hydrates coalesced to form pipes standing on the sediments. Chemical composition and carbon, sulfur, and helium isotopic ratios indicate that the CO(2)-rich fluid was derived from the same magmatic source as dissolved gases in 320 degrees C hydrothermal solution emitted from a nearby black smoker chimney. The CO(2)-rich fluid phase may be separated by subsurface boiling of hydrothermal solutions or by leaching of CO(2)-rich fluid inclusion during posteruption interaction between pore water and volcanogenic sediments.

An adult form of Alexander disease: a novel mutation in glial fibrillary acidic protein.

Glial fibrillary acidic protein (GFAP) mutation has been reported in Alexander disease. We report a patient with the adult form of Alexander disease who shows a novel mutation in GFAP. This case presented with progressive dysarthria, dysphagia and spastic gait on the right side. Brain and spinal cord MRI showed marked atrophy of the medulla oblongata and spinal cord. Abnormal high signal intensities in the ventral medulla oblongata were detected bilaterally. There were no white matter lesions or contrast enhancing lesions. Recently, there have been reports of patients with a juvenile form of Alexander disease presenting with atrophy or signal abnormalities of the medulla or spinal cord. Atrophy of the medulla and spinal cord have specifically been described as suggestive of Alexander disease [1]. Sequence analysis of the GFAP gene of this patient showed a heterozygous c.221T>C mutation, predicting a p.M74T amino acid change. In all patients suspected of Alexander disease on the basis of MRI findings, GFAP analysis is necessary to confirm the diagnosis.

Effects of coronary artery bypass grafting using internal mammary arteries for diabetic patients.

OBJECTIVES: In our institute, internal mammary arteries (IMAs) have been preferred for coronary artery bypass grafting (CABG) in diabetic patients. The purpose of this study was to evaluate the influence of diabetes and IMA grafting on survival after CABG. BACKGROUND: The influence of diabetes on the results of CABG is not well documented, and there is controversy about whether the use of IMAs conveys greater survival benefits to diabetic patients. METHODS: A total of 420 consecutive patients who underwent CABG from April 1990 to July 1998 were reviewed; 211 of these patients had diabetes mellitus at the time of surgery. Internal mammary artery grafts have been used with increasing frequency, and bilateral IMAs have been used when possible since 1993. Internal mammary artery grafts were used in 164 nondiabetic patients (78%) and in 155 diabetic patients (73%). Seventy-eight nondiabetic patients and 74 diabetic patients received bilateral IMA grafts. RESULTS: The postoperative mortality was 2.4% in the nondiabetic and 2.8% in the diabetic group. With regard to postoperative complications, diabetic patients had a significantly higher rate of chest wound infection (p < 0.05), irrespective of whether IMAs were used or not. The use of bilateral IMAs did not increase the risk of chest wound infection in nondiabetic or diabetic patients. Overall survival curve, cardiac death-free curve and cardiac event-free curve were not affected adversely by diabetes, and in diabetic patients, CABG with saphenous veins alone conveyed significantly (p < 0.01) less long-term benefit than did CABG with at least one IMA graft. CONCLUSIONS: It was suggested that IMA grafts should be preferred in diabetic patients.

Comparative study of the affinities of the 5-HT3 receptor antagonists, YM060, YM114 (KAE-393), granisetron and ondansetron in rat vagus nerve and cerebral cortex.

The 5-HT3 receptor blocking properties of YM060, YM114 (KAE-393), granisetron and ondansetron were examined in the vagus nerve and cerebral cortex of rats. 5-HT and 2-methyl-5-HT induced dose-dependent depolarizations of rat isolated vagus nerve with EC50 values of 2.53 (1.93-3.33) x 10(-6) and 4.03 (2.87-5.66) x 10(-6) M, respectively. YM060, YM114 and granisetron dose-dependently antagonized the depolarization of the rat vagus nerve induced by 5-HT, with decreases in the slope and maximal response at higher concentrations. Apparent pA2 values for these antagonists were 10.27 +/- 0.09, 10.12 +/- 0.16 and 9.44 +/- 0.40, respectively. Ondansetron produced a clear rightward shift of the concentration-response curve to 5-HT. The pA2 value was 8.63 (8.23-9.68). YM060 and YM114 at up to 10(-5) M produced no significant depression of the depolarizing responses to DMPP and GABA. YM060, YM114, granisetron and ondansetron displaced specific binding of [3H]GR65630 to rat cortical membranes with pKi values of 10.48 (10.41-10.57), 10.24 (10.18-10.28), 9.15 (9.02-9.28) and 8.70 (8.64-8.77), respectively. An excellent correlation (r = 0.97) was obtained between pA2 values in the vagus nerve and pKi values in the cerebral cortex. YM060, YM114, granisetron and ondansetron showed low affinities for 5-HT1A, 5-HT2 receptor, adrenergic alpha 1, alpha 2, dopamine D2, muscarinic M2, mu-opioid, benzodiazepine and histamine H1 receptors. These results support the possibility that the same type of 5-HT3 receptor occurs in rat vagus nerve and cerebral cortex.

A substance P-containing pathway from the hypothalamic ventromedial nucleus to the medial preoptic area of the rat: an immunohistochemical analysis.

The destruction of the hypothalamic ventromedial nucleus, which contains a group of substance P-like immunoreactive neurons, resulted in a marked ipsilateral reduction of these fibers in the medial preoptic area. To test if and to what extent the substance P-like immunoreactive neurons in the ventromedial nucleus project to the medial preoptic area, we applied a sensitive double-labeling method capable of detecting substance P-like immunoreactivity in neurons retrogradely labeled with biotin-wheat germ agglutinin following injection of the tracer in the medial preoptic area. The appearance of many double-labeled cells in the hypothalamic ventromedial nucleus provides strong evidence for the existence of a prominent substance P containing pathway from the ventromedial nucleus to the medial preoptic area. A few doubled-labeled cells were also seen in the lateral hypothalamus, which therefore seems to be an additional source of substance P-like immunoreactive fibers in the medial preoptic area.

Localization of glycine receptors in the rat central nervous system: an immunocytochemical analysis using monoclonal antibody.

The localization of glycine receptors was immunocytochemically examined in the rat brain using a monoclonal antibody against the affinity-purified glycine receptor. Glycine receptors were concentrated in the lower brainstem, whereas no immunoreactivity was observed in the diencephalon and forebrain except in a few diencephalic nuclei. The highest density of receptors was found in the cranial motor nuclei, reticular formation, parabrachial area, dorsal and ventral cochlear nuclei, and dorsal and ventral tegmental nuclei. Differences were observed in the distribution of immunoreactive elements in the various brain regions. In the cerebellar cortex, the immunoreactivity was exclusively seen along the dendrites of the Purkinje cells. On the other hand, glycine receptors were detected on the cellular membrane of the soma of the cochlear nuclei, trigeminal motor nucleus, parabrachial area, lateral reticular nucleus, dorsal nucleus of the lateral lemniscus, cerebellar nuclei, trigeminal spinal nucleus, anterior horn and reticular formation. In other regions, the receptors were evenly distributed throughout the neuropil.

Distribution of cholinergic neurons and fibers in the hypothalamus of the rat using choline acetyltransferase as a marker.

The distribution of choline-acetyltransferase-like immunoreactive structures in the rat hypothalamus and preoptic area was examined by using avidin-biotin immunocytochemistry. We found that the hypothalamus is richly innervated by the cholinergic neuron system. Sites containing cholinergic neurons of varying density were: medial and lateral preoptic areas, septohypothalamic nucleus, median preoptic area, lateral hypothalamus including the perifornical area, anterior hypothalamic nucleus, arcuate nucleus, dorsomedial hypothalamic nucleus, posterior hypothalamic nucleus, dorsal and ventral premammillary nuclei, neuropil mediodorsal to the anterior hypothalamic nucleus, neuropil ventral to the anterior hypothalamic nucleus and ventromedial hypothalamic nucleus, neuropil between lateral hypothalamus and ventromedial hypothalamus, and neuropil between dorsal premammillary nucleus and posterior hypothalamic nucleus. There were also many varicose and non-varicose fibers in the preoptic area and hypothalamus. Two kinds of varicose fibers, one with strong immunoreactivity and the other with weak immunoreactivity, were seen. Non-varicose fibers were also detected in the optic chiasma and habenulo-interpeduncular tract. These fibers were passing fibers.

Interleukin-6 as a mediator responsible for inflammation-induced increase in plasma angiotensinogen.

The concentration of plasma angiotensinogen increases upon induction of inflammation. Studies were carried out using serum samples collected from mice and rats after injection of lipopolysaccharide (LPS) to determine whether interleukin-6 (IL-6) is a mediator responsible for the inflammation-induced increase of angiotensinogen synthesis in liver cells. Serum collected from mice or rats 2 and 4 hr after injection of LPS contained a factor that stimulated [35S]methionine incorporation into angiotensinogen newly synthesized by rat hepatoma H4IIEC3 (H4) cells. Assay of IL-6 using an IL-6-dependent murine hybridoma, MH60.BSF2 cells, showed the presence of IL-6-like activity in sera of mice or rats 2 and 4 hr after injection of LPS. Anti-mouse IL-6 monoclonal antibody completely inhibited not only the IL-6-like activity present in LPS-treated mouse serum but also the ability of the serum to stimulate angiotensinogen synthesis of H4 cells. These results suggest that increased synthesis of angiotensinogen in the liver after induction of inflammation is mediated by IL-6, a cytokine important in immune reactions and the hepatic acute-phase response.

Localization of neuroactive substances in the rat parabigeminal nucleus: an immunohistochemical study.

The distribution of neurons and fibres that contain substance P, cholecystokinin-8, vasoactive intestinal polypeptide, corticotropin-releasing factor, calcitonin-gene-related peptide, choline acetyltransferase, tyrosine hydroxylase, somatostatin, leucine-enkephalin, and neuropeptide Y was examined in the parabigeminal nucleus of the rat by immunohistochemistry. Many choline acetyltransferase-like immunoreactive or calcitonin-gene-related peptide-like immunoreactive neurons were observed in the dorsal, middle and ventral subdivisions of the parabigeminal nucleus. A few corticotropin-releasing factor-like immunoreactive neurons were also seen in these three subdivisions. The double-immunostaining demonstrated that some choline acetyltransferase-like immunoreactive neurons in the dorsal and ventral subdivisions contained calcitonin-gene-related peptide. Fibres containing cholecystokinin-8, substance P or vasoactive intestinal polypeptide were abundant in the parabigeminal nucleus. Fibres containing cholecystokinin-8 were concentrated in the dorsal and ventral subdivisions, and the lateral margin of the middle subdivision, whereas many fibres containing substance P or vasoactive intestinal polypeptide existed in the lateral half of each subdivision. Fibres containing calcitonin-gene-related peptide or corticotropin-releasing factor were mostly observed around the immunoreactive neurons. Tyrosine hydroxylase-like immunoreactive fibres were scattered in the parabigeminal nucleus.

Afferent reinnervation after lung transplantation in the rat.

Denervation at lung transplantation results in loss of cough reflex and attenuated local defense mechanisms, accounting for increased incidence and severity of infection after lung transplantation. We studied the presence or absence of spontaneous afferent reinnervation in rats at various intervals after orthotopic left pulmonary isografting (n = 52). Normal rats (n = 21) and rats undergoing left hilar stripping (n = 14) served as control subjects. Afferent reinnervation was tested physiologically by reflex bradycardia in response to intravenous infusion of capsaicin (30 microg/kg), an extract of paprika stimulating pulmonary C-fibers. Injection of capsaicin was repeated before and after right pulmonary artery occlusion to divert all pulmonary blood flow to the left lung or isograft. Whereas rats early after surgical denervation lost the reflex after right pulmonary artery occlusion, rats examined 8 months or longer after surgery showed potent reflex bradycardia in response to capsaicin, as did the control rats. Immunohistochemical staining for sensory neuron-specific substances, such as calcitonin gene-related peptide and substance P, were identified only in the right native lung and left pulmonary isografts 2 months or longer after transplantation. Fluorogold was found only in the ipsilateral nodose ganglion after left lung intrapleural injection of the neuron-specific tracer in rats 8 months or longer after denervation. These experiments provide physiological, morphologic, and neurologic evidence suggesting that afferent lung innervation, abolished early after transplantation, is spontaneously reestablished and functioning in the ipsilateral vagus nerve by 8 months after pulmonary isografting in the rat.

Distribution and origin of vasoactive intestinal polypeptide-like immunoreactive fibers in the central amygdaloid nucleus of the rat: an immunocytochemical analysis.

We studied the distribution of vasoactive intestinal polypeptide-like immunoreactive (VIPLI) fibers in the central amygdaloid (AC) nucleus of the rat, using indirect immunofluorescence and the origins of such fibers using a combination of retrograde tracing with immunocytochemistry. VIPLI fibers formed a dense fiber plexus in the lateral subdivision of the AC nucleus, but other subdivisions showed little immunoreactivity. Destruction of the supramammillary (SuM) region and the adjacent lateral hypothalamus, both of which contained a group of VIPLI neurons, resulted in the marked reduction of VIPLI fibers in the ipsilateral AC nucleus, indicating that many of the fibers in the AC nucleus originate from these two areas. This assumption was supported by the finding that injection of fast blue dye into the AC nucleus labeled the VIPLI neurons in the SuM region and lateral hypothalamus.

Distribution of corticotropin-releasing factor and calcitonin gene-related peptide in the developing mouse cerebellum.

Corticotropin-releasing factor (CRF)-like immunoreactive (IR) fibers were investigated ontogenically in the mouse cerebellum. CRF-IR was detected in the climbing fiber and mossy fibers as in other species. In addition, CRF-IR dense fiber plexuses were detected from postnatal day (PD) 2 to 9, in the developing Purkinje cell layer of the vermal lobules, paraflocculus, flocculus and crus 1 ansiform lobule, gradually forming a pericellular nest around the Purkinje cell somata. Immunoelectron-microscopical analysis showed that dense fibers made synaptic contacts with the Purkinje cell somata on PD 7. In the lobules mentioned above, CRF-IR dense fibers showed parasagittal banded patterns. Calcitonin gene-related peptide (CGRP)-IR showed similar fiber bands at these stages. Interestingly, these two patterns of peptidergic fiber bands were complementary in distribution. From around PD 9, CRF-IR fibers lost the immunoreactive dots in the Purkinje cell layer. Immunoreactivity at this stage was observed in the axons projecting to the molecular layer, and thin CRF-IR fibers began to appear in the neighboring area. Numerous typical climbing fiber-like CRF-IR fibers were found throughout the cerebellar cortex from PD 16 to adult. The inferior olivary complex (the origin of climbing fibers) appears to be the origin of these dense fiber plexuses as CRF-IR cells were already present from PD 2 in the dorsal cap nucleus, beta subnucleus and caudomedial part of the accessory olivary nucleus. No neurons containing both CRF and CGRP immunoreactivities were observed. These results suggest that CGRP- and CRF-IR developing climbing fibers innervate different compartments of Purkinje cells, especially in the vestibular cerebellar cortex in mice. Furthermore, CRF-IR fibers gradually changed to become typical climbing fibers, while CGRP-IR disappeared altogether.

Stroke after coronary artery bypass grafting in patients with cerebrovascular disease.

BACKGROUND: Stroke has been associated with a significantly increased mortality from coronary artery bypass grafting (CABG). To determine the predictors of stroke in patients undergoing CABG, we collected data on 472 consecutive patients. METHODS: From March 1991 to March 1999, all patients undergoing CABG at our institution underwent routine duplex scanning of the extracranial carotid and vertebral arteries. Seven patients with symptomatic carotid stenosis were treated by carotid endarterectomy (CEA) before CABG. RESULTS: There was a 10-fold increase in mortality (12.5%) associated with postoperative stroke. Many variables were analyzed by a multivariate technique and the severity of extracranial carotid artery stenosis was determined to be the only independent predictor of postoperative stroke (p < 0.01). None of the patients with carotid artery occlusion and none of the patients who underwent CEA before CABG experienced a stroke. CONCLUSIONS: To reduce the stroke rate, the indications for prophylactic CEA may be extended for asymptomatic patients with carotid artery stenosis greater than 75%.

Afferent and efferent enkephalinergic systems of the tegmental nuclei of Gudden in the rat: an immunocytochemical study.

We studied the afferent source of L-enkephalin-like immunoreactive (L-ENKLI) fibers in the ventral tegmental nucleus (VT) of Gudden, and efferent and afferent connections of L-ENKLI structures in the dorsal part of the dorsal tegmental nucleus (DDT) of Gudden in the rat, using immunocytochemistry combined with knife-cut and lesion experiments. The VT had a dense plexus of L-ENKLI fibers but no L-ENKLI cells. Destruction of the dorsal premammillary nucleus and medial mammillary nucleus pars medialis, which contained a large group of L-ENKLI neurons, markedly reduced L-ENKLI fibers in the ipsilateral VT, suggesting that most of these fibers originate in these nuclei. The DDT contained a large collection of L-ENKLI neurons together with dense L-ENKLI fibers. Destruction of the DDT caused a contralateral marked reduction of L-ENKLI fibers in the dorsolateral part of the interpeduncular nucleus (DL), suggesting that L-ENKLI neurons in the DDT project contralaterally to the DL. L-ENKLI fibers in the DDT may be of intrinsic origin.

An enkephalinergic projection from the hypothalamic paraventricular nucleus to the hypothalamic ventromedial nucleus of the rat: an experimental immunohistochemical study.

The distribution and origins of enkephalin-like immunoreactive (ENK-IR) fibers in the nucleus ventromedialis hypothalami (vm) of the rat were examined using immunohistochemistry. A dense plexus was evenly distributed in the vm with no regional differences. A group of ENK-IR neurons was concentrated in the ventrolateral portion of the vm. The destruction of the hypothalamic paraventricular nucleus (pv) which contained numerous ENK-IR neurons, resulted in a marked decrease in ENK-IR fibers in the vm on the operated side. In addition, the destruction of the magnocellular portion of the pv, while leaving most of parvocellular portion intact, failed to substantially decrease the ENK-IR fibers in the vm, suggesting that these fibers originate from ENK-IR neurons located in the parvocellular portion of the pv. The present study further showed that the axons of these neurons first proceeded laterally to the perifornical area, next ran ventrolaterally to the ventrolateral portion of the anterior hypothalamic nucleus and finally turned medially to the vm.

Coexistence of acetylcholine and calcitonin gene-related peptide in the vestibular efferent neurons in the rat.

An immunocytochemical study combined with the retrograde tracer technique was performed in the rat to assess the vestibular efferent system. All 3 neuron groups which give rise to axons terminating on the vestibular end-organs were cholinergic, i.e., a group dorsolateral to the genu of the facial nerve (DL), a group dorsomedial to this genu (M), and scattered cells in the parvocellular reticular nucleus (PCRt). In addition, we further demonstrated that about 55% of the cholinergic cells in DL had calcitonin gene-related peptide.

Presence of VIP fibers of sensory origin in the rat trachea.

The presence of vasoactive intestinal peptide (VIP) immunoreactive nerve fibers (LI) in the respiratory tract of mammals is well documented. These fibers are known to originate from parasympathetic postganglionic neurons and to be associated with blood vessels, submucosal glands, and with smooth muscle. We found that, in addition to this, the epithelial layer of the rat trachea also contains VIP-LI fibers. Vagotomy or ligation of the cervical portion of the vagus nerve resulted respectively in a decrease of VIP-LI fibers within the epithelium or in the accumulation of VIP in axons proximal to the site of transection or ligation, whereas no changes were seen in other parts of the trachea. On the other hand, capsaicin pretreatment also caused similar changes to the surgical procedures. These findings indicate that VIP-LI fibers in the tracheal epithelium of the rat are supplied by the sensory ganglia of the vagus nerve.

Ultrastructural evidence of direct synaptic contact of catecholamine terminals with oxytocin-containing neurons in the parvocellular portion of the rat hypothalamic paraventricular nucleus.

Ultrastructural evidence of the direct interaction between catecholamine (CA) terminals and oxytocin (OX)-containing neurons in the parvocellular portion of the rat hypothalamic paraventricular nucleus was demonstrated using immunohistochemistry combined with false transmitter (5-hydroxydopamine, 5-OHDA) histochemistry. At the parvocellular portion, 5-OHDA labelled CA terminals make synaptic contact with proximal dendrites or somas of OX-positive cells, suggesting that the ascending CA system monosynaptically regulates the extrahypothalamic OX system.

Somatostatinergic neurons in the insular cortex project to the spinal cord: combined retrograde axonal transport and immunohistochemical study.

A double-labeling method combining immunohistochemistry and a retrograde tracer technique using biotin-horseradish peroxidase (B-HRP) was employed to identify a descending somatostatinergic fiber system from the insular cortex to the spinal cord. Injection of B-HRP into the spinal cord at cervical or lumbar levels resulted in the labeling of a number of neurons in the insular cortex. Simultaneous immunostaining revealed the existence of double-labeled neurons in the insular cortex. The result provides direct evidence for the presence of a descending somatostatinergic pathway from the insular cortex to lumbar levels of the spinal cord.

Distribution and origins of neurotensin-containing fibers in the nucleus ventromedialis hypothalami of the rat: an experimental immunohistochemical study.

The distribution and origins of neurotensin (NT)-containing fibers in the nucleus ventromedialis hypothalami (VM) of the rat were investigated experimentally using an indirect immunofluorescence technique. A dense plexus of NT-like immunoreactive (NTI) fibers which was composed of very fine varicosities was identified in the VM. Although they were distributed throughout its entire rostrocaudal extent, the distribution was uneven. The highest density was identified in the dorsomedial part of the VM. In the central part, a less numerous but still moderate number of NTI fibers was detected in its dorsal part. But in a ventrolateral direction, they decreased in number and in the ventrolateral part only a few NTI fibers were seen. The present study demonstrated experimentally that these fibers originate from the medial nucleus of the amygdaloid complex (AM), since destruction of the AM resulted in a marked reduction of NTI fibers ipsilaterally in the VM. These findings suggest that the AM influences the VM's functions via neurotensin-like immunoreactive fibers.